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Rat NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
ELK1693
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48 T $320.00
96 T $458.00
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Overview

Product name: Rat NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
Reactivity: Rat
Alternative Names: NFKB3; p65; REL-A; Nuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B-Cells 3; Transcription Factor P65; Nuclear factor NF-kappa-B p65 subunit; RELA; V-Rel Reticuloendotheliosis Viral Oncogene Homolog A
Assay Type: Sandwich
Sensitivity: 0.058 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: Tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Apoptosis;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Rat NFκB-p65. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat NFκB-p65. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Rat NFκB-p65, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rat NFκB-p65 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
10.00 1.978 1.891
5.00 1.536 1.449
2.50 1.184 1.097
1.25 0.841 0.754
0.63 0.554 0.467
0.32 0.381 0.294
0.16 0.190 0.103
0.00 0.087 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant NFκB-p65 and the recovery rates were calculated by comparing the measured value to the expected amount of NFκB-p65 in samples.
Matrix Recovery range Average
serum(n=5) 83-97% 90%
EDTA plasma(n=5) 81-95% 88%
Heparin plasma(n=5) 85-95% 90%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NFκB-p65 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 86-94% 93-102% 89-97% 85-94%
EDTA plasma(n=5) 91-99% 83-97% 87-105% 87-99%
Heparin plasma(n=5) 85-96% 87-101% 89-102% 85-98%